Cytokine-Induced Guanylate Binding Protein 1 (GBP1) Release from Human Ovarian Cancer Cells

We showed that IL-27 shares several effects with IFN-γ in human cancer cells. To identify novel extracellular mediators, potentially involved in epithelial ovarian cancer (EOC) biology, we analyzed the effect of IL-27 or IFN-γ on the secretome of cultured EOC cells by mass-spectrom...

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Main Authors: Grazia Carbotti, Andrea Petretto, Elisabeth Naschberger, Michael Stürzl, Stefania Martini, Maria Cristina Mingari, Gilberto Filaci, Silvano Ferrini, Marina Fabbi
Format: Article
Language:English
Published: MDPI AG 2020-02-01
Series:Cancers
Subjects:
Online Access:https://www.mdpi.com/2072-6694/12/2/488
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spelling doaj-5b865f2a8c40482c9ef8ddfc01ac4f2d2020-11-25T01:42:27ZengMDPI AGCancers2072-66942020-02-0112248810.3390/cancers12020488cancers12020488Cytokine-Induced Guanylate Binding Protein 1 (GBP1) Release from Human Ovarian Cancer CellsGrazia Carbotti0Andrea Petretto1Elisabeth Naschberger2Michael Stürzl3Stefania Martini4Maria Cristina Mingari5Gilberto Filaci6Silvano Ferrini7Marina Fabbi8IRCCS Ospedale Policlinico San Martino, Biotherapies Unit, Largo R. Benzi 10, 16132 Genoa, ItalyCore Facilities—Clinical Proteomics and Metabolomics, IRCCS Istituto Giannina Gaslini, Via Gerolamo Gaslini 5, 16147 Genoa, ItalyDivision of Molecular and Experimental Surgery, University Medical Center Erlangen, Friedrich-Alexander-University Erlangen-Nürnberg, Schwabachanlage 12, 91054 Erlangen, GermanyDivision of Molecular and Experimental Surgery, University Medical Center Erlangen, Friedrich-Alexander-University Erlangen-Nürnberg, Schwabachanlage 12, 91054 Erlangen, GermanyIRCCS Ospedale Policlinico San Martino, Immunology Unit, Largo R. Benzi 10, 16132 Genoa, ItalyIRCCS Ospedale Policlinico San Martino, Immunology Unit, Largo R. Benzi 10, 16132 Genoa, ItalyIRCCS Ospedale Policlinico San Martino, Biotherapies Unit, Largo R. Benzi 10, 16132 Genoa, ItalyIRCCS Ospedale Policlinico San Martino, Biotherapies Unit, Largo R. Benzi 10, 16132 Genoa, ItalyIRCCS Ospedale Policlinico San Martino, Biotherapies Unit, Largo R. Benzi 10, 16132 Genoa, ItalyWe showed that IL-27 shares several effects with IFN-γ in human cancer cells. To identify novel extracellular mediators, potentially involved in epithelial ovarian cancer (EOC) biology, we analyzed the effect of IL-27 or IFN-γ on the secretome of cultured EOC cells by mass-spectrometry (nano-UHPLC-MS/MS). IL-27 and IFN-γ modulate the release of a limited fraction of proteins among those induced in the whole cell. We focused our attention on GBP1, a guanylate-binding protein and GTPase, which mediates several biological activities of IFNs. Cytokine treatment induced GBP1, 2, and 5 expressions in EOC cells, but only GBP1 was secreted. ELISA and immunoblotting showed that cytokine-stimulated EOC cells release full-length GBP1 in vitro, through non-classical pathways, not involving microvesicles. Importantly, full-length GBP1 accumulates in the ascites of most EOC patients and ex-vivo EOC cells show constitutive tyrosine-phosphorylated STAT1/3 proteins and GBP1 expression, supporting a role for Signal Transducer And Activator Of Transcription (STAT)-activating cytokines in vivo. High GBP1 gene expression correlates with better overall survival in the TCGA (The Cancer Genome Atlas) dataset of EOC. In addition, GBP1 transfection partially reduced EOC cell viability in an MTT assay. Our data show for the first time that cytokine-stimulated tumor cells release soluble GBP1 in vitro and in vivo and suggest that GBP1 may have anti-tumor effects in EOC.https://www.mdpi.com/2072-6694/12/2/488ovarian cancerinterleukin-27interferon-γsecretomeguanylate binding protein 1microenvironment
collection DOAJ
language English
format Article
sources DOAJ
author Grazia Carbotti
Andrea Petretto
Elisabeth Naschberger
Michael Stürzl
Stefania Martini
Maria Cristina Mingari
Gilberto Filaci
Silvano Ferrini
Marina Fabbi
spellingShingle Grazia Carbotti
Andrea Petretto
Elisabeth Naschberger
Michael Stürzl
Stefania Martini
Maria Cristina Mingari
Gilberto Filaci
Silvano Ferrini
Marina Fabbi
Cytokine-Induced Guanylate Binding Protein 1 (GBP1) Release from Human Ovarian Cancer Cells
Cancers
ovarian cancer
interleukin-27
interferon-γ
secretome
guanylate binding protein 1
microenvironment
author_facet Grazia Carbotti
Andrea Petretto
Elisabeth Naschberger
Michael Stürzl
Stefania Martini
Maria Cristina Mingari
Gilberto Filaci
Silvano Ferrini
Marina Fabbi
author_sort Grazia Carbotti
title Cytokine-Induced Guanylate Binding Protein 1 (GBP1) Release from Human Ovarian Cancer Cells
title_short Cytokine-Induced Guanylate Binding Protein 1 (GBP1) Release from Human Ovarian Cancer Cells
title_full Cytokine-Induced Guanylate Binding Protein 1 (GBP1) Release from Human Ovarian Cancer Cells
title_fullStr Cytokine-Induced Guanylate Binding Protein 1 (GBP1) Release from Human Ovarian Cancer Cells
title_full_unstemmed Cytokine-Induced Guanylate Binding Protein 1 (GBP1) Release from Human Ovarian Cancer Cells
title_sort cytokine-induced guanylate binding protein 1 (gbp1) release from human ovarian cancer cells
publisher MDPI AG
series Cancers
issn 2072-6694
publishDate 2020-02-01
description We showed that IL-27 shares several effects with IFN-γ in human cancer cells. To identify novel extracellular mediators, potentially involved in epithelial ovarian cancer (EOC) biology, we analyzed the effect of IL-27 or IFN-γ on the secretome of cultured EOC cells by mass-spectrometry (nano-UHPLC-MS/MS). IL-27 and IFN-γ modulate the release of a limited fraction of proteins among those induced in the whole cell. We focused our attention on GBP1, a guanylate-binding protein and GTPase, which mediates several biological activities of IFNs. Cytokine treatment induced GBP1, 2, and 5 expressions in EOC cells, but only GBP1 was secreted. ELISA and immunoblotting showed that cytokine-stimulated EOC cells release full-length GBP1 in vitro, through non-classical pathways, not involving microvesicles. Importantly, full-length GBP1 accumulates in the ascites of most EOC patients and ex-vivo EOC cells show constitutive tyrosine-phosphorylated STAT1/3 proteins and GBP1 expression, supporting a role for Signal Transducer And Activator Of Transcription (STAT)-activating cytokines in vivo. High GBP1 gene expression correlates with better overall survival in the TCGA (The Cancer Genome Atlas) dataset of EOC. In addition, GBP1 transfection partially reduced EOC cell viability in an MTT assay. Our data show for the first time that cytokine-stimulated tumor cells release soluble GBP1 in vitro and in vivo and suggest that GBP1 may have anti-tumor effects in EOC.
topic ovarian cancer
interleukin-27
interferon-γ
secretome
guanylate binding protein 1
microenvironment
url https://www.mdpi.com/2072-6694/12/2/488
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